Re: [DIYbio] Plasmid miniprep kit

Wow, Cory, that's great! I look forward to reading and trying that.

On Sunday, 25 March 2012 20:46:10 UTC-7, cory....@gmail.com wrote:

No autoclaving necessary.

Also, I wrote an article for the next volume of Citizen Science
Quarterly, which should be out shortly I think, that has recipes for
all the miniprep solutions using only household ingredients.  I found
that sodium chloride works just fine in place of guanidine
hydrochloride as long as the concentration of salt in the final
mixture (buffer P1 + P2 + N3) is above 2M and the pH is below 5.5.  My
homemade recipe for N3 is 11.5g table salt, 3.7g potassium chloride,
and 43mL of distilled white vinegar (no extra water added).  You can
get the potassium chloride at the grocery store (I found it at
Ralphs).  It's sold by Morton Salt as some sort of additive for water
softener tanks.  Anyways, if you give it a try let me know how it
works.

Also, an added benefit of using the NaCl is that very little RNA is
bound to the silica compared to guanidine (which is why kits come with
RNase to add to buffer P1).  There is a little RNA that comes along
for the ride, but I imagine if you play around the the pH and salt
concentration you can probably minimize that even more.

-cory

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